Epithelial-to-mesenchymal transition drives a pro-metastatic Golgi compaction process through scaffolding protein PAQR11
Xiaochao Tan, Priyam Banerjee, Hou-Fu Guo, Stephen Ireland, Daniela Pankova, Young-ho Ahn, Irodotos Michail Nikolaidis, Xin Liu, Yanbin Zhao, Yongming Xue, Alan R. Burns, Jonathon Roybal, Don L. Gibbons, Tomasz Zal, Chad J. Creighton, Daniel Ungar, Yanzhuang Wang, Jonathan M. Kurie
Xiaochao Tan, Priyam Banerjee, Hou-Fu Guo, Stephen Ireland, Daniela Pankova, Young-ho Ahn, Irodotos Michail Nikolaidis, Xin Liu, Yanbin Zhao, Yongming Xue, Alan R. Burns, Jonathon Roybal, Don L. Gibbons, Tomasz Zal, Chad J. Creighton, Daniel Ungar, Yanzhuang Wang, Jonathan M. Kurie
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Research Article Cell biology Oncology

Epithelial-to-mesenchymal transition drives a pro-metastatic Golgi compaction process through scaffolding protein PAQR11

Abstract

Tumor cells gain metastatic capacity through a Golgi phosphoprotein 3–dependent (GOLPH3-dependent) Golgi membrane dispersal process that drives the budding and transport of secretory vesicles. Whether Golgi dispersal underlies the pro-metastatic vesicular trafficking that is associated with epithelial-to-mesenchymal transition (EMT) remains unclear. Here, we have shown that, rather than causing Golgi dispersal, EMT led to the formation of compact Golgi organelles with improved ribbon linking and cisternal stacking. Ectopic expression of the EMT-activating transcription factor ZEB1 stimulated Golgi compaction and relieved microRNA-mediated repression of the Golgi scaffolding protein PAQR11. Depletion of PAQR11 dispersed Golgi organelles and impaired anterograde vesicle transport to the plasma membrane as well as retrograde vesicle tethering to the Golgi. The N-terminal scaffolding domain of PAQR11 was associated with key regulators of Golgi compaction and vesicle transport in pull-down assays and was required to reconstitute Golgi compaction in PAQR11-deficient tumor cells. Finally, high PAQR11 levels were correlated with EMT and shorter survival in human cancers, and PAQR11 was found to be essential for tumor cell migration and metastasis in EMT-driven lung adenocarcinoma models. We conclude that EMT initiates a PAQR11-mediated Golgi compaction process that drives metastasis.

Authors

Xiaochao Tan, Priyam Banerjee, Hou-Fu Guo, Stephen Ireland, Daniela Pankova, Young-ho Ahn, Irodotos Michail Nikolaidis, Xin Liu, Yanbin Zhao, Yongming Xue, Alan R. Burns, Jonathon Roybal, Don L. Gibbons, Tomasz Zal, Chad J. Creighton, Daniel Ungar, Yanzhuang Wang, Jonathan M. Kurie

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Figure 8

High PAQR11 levels enhance lung adenocarcinoma metastatic properties.

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High PAQR11 levels enhance lung adenocarcinoma metastatic properties. (A...
(A) Kaplan-Meier plots of compendia of lung adenocarcinoma cohorts (left) and over 30 tumor types (right). The groups with highest (“top third”), lowest (“bottom third”), and intermediate (“middle third”) PAQR11 mRNA levels were compared. Numbers of patients alive at each time point indicated under graphs. P values determined by univariate Cox and stratified log-rank test correcting for cancer type. (B) Scatter plots of primary tumor weights (left) and lung metastasis numbers (right) per mouse (dots). (C) Bright-field micrographs of cells at the leading edges of scratch-wounded cultures of 393P_ZEB1 cells stably transfected with scrambled control shRNA (shCTL) or PAQR11 shRNA (shPAQR11). Bar graph shows percent wound closure 24 and 48 hours after placing of wounds. (D) Time-lapse tracing of movements of 344SQ cells stably transfected with scrambled control shRNA (shCTL) or PAQR11 shRNA (shPAQR11). Movements of individual cells over 6 hours indicated by colored lines (n = 15 cells per group). Bar graphs show quantification of cell speed (left) and directionality (right). (E) Bright-field micrograph of a 393P_ZEB1 spheroid in collagen showing cells invading singly (red arrow) or collectively (white arrow). Scale bar: 100 μm. The bar and whisker plot shows quantification of single-cell (single) and collective invasions (n > 20 spheroids per group). (F) Bright-field micrographs of migrated and invaded cells in Boyden chambers. 393P_ZEB1 cells stably transfected with scrambled control shRNA (shCTL) or 1 of 2 distinct PAQR11 shRNAs (shPAQR11#1 or shPAQR11#3) stained with crystal violet (purple). Scale bar: 200 μm. Bar graph shows quantification of migrated and invaded cells. Unless otherwise indicated, results represent averages of triplicate samples per condition. P values were determined using 2-tailed Student’s t test or ANOVA for comparisons between 2 groups or more than 2 groups, respectively. Results were replicated (n ≥ 2 experiments).